Related Experiment Video
Updated: Aug 17, 2026

Optimized Analysis of DNA Methylation and Gene Expression from Small, Anatomically-defined Areas of the Brain
Published on: July 12, 2012
Picogram cloning and direct in situ sequencing of DNA from gel pieces
E Meijerink1, B Kozulic, G Stranzinger
1Cytos Biotechnology AG, Schlieren, Switzerland.
Abstract:
We describe a simple and rapid procedure for cloning and sequencing of DNA fragments separated by gel electrophoresis, using novel hydrophilic gels, Clearose BG, Spreadex, and Poly(NAT), that do not melt at 95 degrees C. For cloning, a band of interest is excised precisely and incubated in an extraction buffer containing 5-10 mM MgCl2 at 70 degrees C for 15-45 min. The eluted DNA is added directly to the plasmid solution. Using a topoisomerase-based ligation system, we were able to transform bacteria with a few picograms of DNA and isolate recombinant clones. For in situ sequencing, the DNA in the gel serves as the template. No treatment before cycle sequencing is necessary for fragments up to 500 bp.
Related Concept Videos
DNA Isolation
DNA Agarose Gel Electrophoresis
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Maxam-Gilbert Sequencing
Challenges of the Maxam-Gilbert Method
The...

